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Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: Afatinib and Temozolomide combination inhibits tumorigenesis by targeting EGFRvIII-cMet signaling in glioblastoma cells

Fig. 1

Afatinib and TMZ combination differentially inhibit proliferation and clonogenic survival of EGFR and EGFRvIII expressing GBM cells. a, b EGFR and EGFRvIII expression differentially effect cell proliferation. a U87MG, U87EGFR WT, U87EGFRvIII, U87EGFRvIII DK, U251, U251EGFR WT (Tet-inducible system) and U251EGFRvIII (Tet-inducible system) cell lysates were analyzed for EGFR (full length), EGFRvIII and pEGFR (Tyr-1068). b The graph shows increased proliferation rate of U87EGFRvIII cells compared to U87MG, U87EGFR WT, and U87EGFRvIII DK cells. Tet - tetracycline. c, d U87MG, U87EGFR WT, U87EGFRvIII and U87EGFRvIII DK cells were seeded and treated as specified for 48 h. After washing with PBS, cells were allowed to grow for 12 days in 2% media. Colonies formed were fixed with methanol, stained with crystal violet solution, dissolved in 10% acetic acid and absorbance was measured at 570 nm. The graph shows the mean (± SD) percentage of colony formation. The experiment was repeated three times (*$ P ≤ 0.05); * significant compared to control; $ significant compared to TMZ. e, f Afatinib and TMZ combination decreases the anchorage-independent growth of TMZ-resistant EGFRvIII GBM cells. U87MG, U87 EGFR WT, U87EGFRvIII and U87EGFRvIII DK cells (5 × 103) were seeded with 0.25% agarose on the top of the 0.5% base agar. After overnight incubation, cells were incubated with TMZ (25 μM) or afatinib (1 μM) or combination of both for a month. The 0.1% crystal violet stained colonies were counted (n = 3). The graph shows mean (± SD) number of small (blue) and big colonies (orange). (* P ≤ 0.05); * number of small colonies, significant compared to control; # number of big colonies, significant compared to control

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